Development of a toolkit for piggyBac-mediated integrative transfection of the human filarial parasite Brugia malayi.
BACKGROUND:The human filarial parasites cause diseases that are among the most important causes of morbidity in the developing world. The elimination programs targeting these infections rely on a limited number of drugs, making the identification of new chemotherapeutic agents a high priority. The s...
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ftdoajarticles:oai:doaj.org/article:9b4613c7bb45473485144410bd81365e 2023-05-15T15:07:14+02:00 Development of a toolkit for piggyBac-mediated integrative transfection of the human filarial parasite Brugia malayi. Canhui Liu Amruta S Mhashilkar Johan Chabanon Shulin Xu Sara Lustigman John H Adams Thomas R Unnasch 2018-05-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0006509 https://doaj.org/article/9b4613c7bb45473485144410bd81365e EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC5983866?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0006509 https://doaj.org/article/9b4613c7bb45473485144410bd81365e PLoS Neglected Tropical Diseases, Vol 12, Iss 5, p e0006509 (2018) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2018 ftdoajarticles https://doi.org/10.1371/journal.pntd.0006509 2022-12-31T00:41:21Z BACKGROUND:The human filarial parasites cause diseases that are among the most important causes of morbidity in the developing world. The elimination programs targeting these infections rely on a limited number of drugs, making the identification of new chemotherapeutic agents a high priority. The study of these parasites has lagged due to the lack of reverse genetic methods. METHODOLOGY/PRINCIPAL FINDINGS:We report a novel co-culture method that results in developmentally competent infective larvae of one of the human filarial parasites (Brugia malayi) and describe a method to efficiently transfect the larval stages of this parasite. We describe the production of constructs that result in integrative transfection using the piggyBac transposon system, and a selectable marker that can be used to identify transgenic parasites. We describe the production and use of dual reporter plasmids containing both a secreted luciferase selectable marker and fluorescent protein reporters that will be useful to study temporal and spatial patterns of gene expression. CONCLUSIONS/SIGNIFICANCE:The methods and constructs reported here will permit the efficient production of integrated transgenic filarial parasite lines, allowing reverse genetic technologies to be applied to all life cycle stages of the parasite. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 12 5 e0006509 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
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English |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Canhui Liu Amruta S Mhashilkar Johan Chabanon Shulin Xu Sara Lustigman John H Adams Thomas R Unnasch Development of a toolkit for piggyBac-mediated integrative transfection of the human filarial parasite Brugia malayi. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
BACKGROUND:The human filarial parasites cause diseases that are among the most important causes of morbidity in the developing world. The elimination programs targeting these infections rely on a limited number of drugs, making the identification of new chemotherapeutic agents a high priority. The study of these parasites has lagged due to the lack of reverse genetic methods. METHODOLOGY/PRINCIPAL FINDINGS:We report a novel co-culture method that results in developmentally competent infective larvae of one of the human filarial parasites (Brugia malayi) and describe a method to efficiently transfect the larval stages of this parasite. We describe the production of constructs that result in integrative transfection using the piggyBac transposon system, and a selectable marker that can be used to identify transgenic parasites. We describe the production and use of dual reporter plasmids containing both a secreted luciferase selectable marker and fluorescent protein reporters that will be useful to study temporal and spatial patterns of gene expression. CONCLUSIONS/SIGNIFICANCE:The methods and constructs reported here will permit the efficient production of integrated transgenic filarial parasite lines, allowing reverse genetic technologies to be applied to all life cycle stages of the parasite. |
format |
Article in Journal/Newspaper |
author |
Canhui Liu Amruta S Mhashilkar Johan Chabanon Shulin Xu Sara Lustigman John H Adams Thomas R Unnasch |
author_facet |
Canhui Liu Amruta S Mhashilkar Johan Chabanon Shulin Xu Sara Lustigman John H Adams Thomas R Unnasch |
author_sort |
Canhui Liu |
title |
Development of a toolkit for piggyBac-mediated integrative transfection of the human filarial parasite Brugia malayi. |
title_short |
Development of a toolkit for piggyBac-mediated integrative transfection of the human filarial parasite Brugia malayi. |
title_full |
Development of a toolkit for piggyBac-mediated integrative transfection of the human filarial parasite Brugia malayi. |
title_fullStr |
Development of a toolkit for piggyBac-mediated integrative transfection of the human filarial parasite Brugia malayi. |
title_full_unstemmed |
Development of a toolkit for piggyBac-mediated integrative transfection of the human filarial parasite Brugia malayi. |
title_sort |
development of a toolkit for piggybac-mediated integrative transfection of the human filarial parasite brugia malayi. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2018 |
url |
https://doi.org/10.1371/journal.pntd.0006509 https://doaj.org/article/9b4613c7bb45473485144410bd81365e |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 12, Iss 5, p e0006509 (2018) |
op_relation |
http://europepmc.org/articles/PMC5983866?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0006509 https://doaj.org/article/9b4613c7bb45473485144410bd81365e |
op_doi |
https://doi.org/10.1371/journal.pntd.0006509 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
12 |
container_issue |
5 |
container_start_page |
e0006509 |
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1766338779648360448 |